公开(公告)号：WO2023086670A2

公开(公告)日：2023-05-19

申请号：PCT/US2022/049950

申请日：2022-11-15

Applicant: THE TRUSTEES OF PRINCETON UNIVERSITY

Inventor： MYHRVOLD, Cameron ,  KIM, Gaeun ,  ARIZTI-SANZ, Jon ,  MCNULTY, Ryan

IPC: C12Q1/689

Abstract: Described herein is a method of screening for a nuclease with an altered nuclease activity comprising: a) forming a first compartmentalizing reaction comprising a carrier and a variant nuclease template nucleic acid comprising a coding region for a variant nuclease, and amplifying said coding region for said variant nuclease, to obtain variant nuclease encoding nucleic acid; b) forming a second compartmentalizing reaction comprising said variant nuclease encoding nucleic acid, and performing an in vitro transcription and translation reaction to obtain variant nuclease polypeptides; and c) forming a third compartmentalizing reaction comprising said variant nuclease polypeptides, and assaying said variant nuclease polypeptides for the altered nuclease activity of said nuclease.